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1.
Comparison of canine and feline adipose-derived mesenchymal stem cells/medicinal signaling cells with regard to cell surface marker expression, viability, proliferation, and differentiation potential
Metka Voga, Valerija Kovač, Gregor Majdič, 2021, izvirni znanstveni članek

Opis: Remarkable immunomodulatory abilities of mesenchymal stem cells, also called multipotent mesenchymal stromal cells or medicinal signaling cells (MSCs), have entailed significant advances in veterinary regenerative medicine in recent years. Despite positive outcomes from MSC therapies in various diseases in dogs and cats, differences in MSC characteristics between small animal veterinary patients are not well-known. We performed a comparative study of cells' surface marker expression, viability, proliferation, and differentiation capacity of adipose-derived MSCs (ADMSCs) from dogs and domestic cats. The same growth media and methods were used to isolate, characterize, and culture canine and feline ADMSCs. Adipose tissue was collected from 11 dogs and 8 cats of both sexes. The expression of surface markers CD44, CD90, and CD34 was detected by flow cytometry. Viability at passage 3 was measured with the hemocytometer and compared to the viability measured by flow cytometry after 1 day of handling. The proliferation potential of MSCs was measured by calculating cell doubling and cell doubling time from second to eighth passage. Differentiation potential was determined at early and late passages by inducing cells toward adipogenic, osteogenic, and chondrogenic differentiation using commercial media. Our study shows that the percentage of CD44+CD90+ and CD34−/− cells is higher in cells from dogs than in cells from cats. The viability of cells measured by two different methods at passage 3 differed between the species, and finally, canine ADMSCs possess greater proliferation and differentiation potential in comparison to the feline ADMSCs.
Ključne besede: mesenchymal stem cells, dog, cat, comparison, proliferation, differentiation, cell surface marker, viability
Objavljeno v DKUM: 21.10.2024; Ogledov: 0; Prenosov: 2
.pdf Celotno besedilo (2,46 MB)
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2.
Behaviour of cellular metamaterials under impact loading conditions : doctoral disertation
Yunus Emre Yilmaz, 2024, doktorska disertacija

Opis: This doctoral thesis investigates cell-size-graded metallic and non-metallic Triply Periodic Minimal Surface (TPMS) structures' behavior under varying loading rates. Using experimental tests, analytical calculations, and advanced computer simulations, the research explores the interplay between material properties, cell size grading, and deformation mechanisms under different strain rates. The study focuses on enhancing the Direct Impact Hopkinson Bar (DIHB) setup for accurate force and displacement measurements and pioneering a method for quantifying inertial forces, critical at high strain rates. Key findings show that cell-size grading significantly affects deformation patterns, with initial deformation occurring in regions with smaller and lower stiffness cells across different loading rates and TPMS geometries. The research also highlights topology's influence on mechanical response, with photopolymer-based diamond structures showing superior energy absorption and gas-atomized steel structures favoring gyroid configurations. This underscores the importance of considering both topology and base material selection during TPMS design. The study demonstrates the increasing prominence of inertial forces as deformation rates rise, impacting structural response and failure likelihood in TPMS structures. These insights inform the design of optimized cellular metamaterials for high-performance applications requiring superior energy absorption and structural integrity under high loading rates. The research advances material characterization techniques and computational modelling capabilities, contributing to the development of next-generation cellular metamaterials for broader engineering applications.
Ključne besede: Triply Periodic Minimal Surfaces, TPMS, Cell-size-grading, Impact, High-strainrate, Digital Image Correlation
Objavljeno v DKUM: 17.10.2024; Ogledov: 0; Prenosov: 16
.pdf Celotno besedilo (46,48 MB)

3.
Dual mode of action of acetylcholine on cytosolic calcium oscillations in pancreatic beta and acinar cells in situ
Nastja Sluga, Sandra Postić, Srdjan Sarikas, Ya-Chi Huang, Andraž Stožer, Marjan Rupnik, 2021, izvirni znanstveni članek

Opis: Cholinergic innervation in the pancreas controls both the release of digestive enzymes to support the intestinal digestion and absorption, as well as insulin release to promote nutrient use in the cells of the body. The effects of muscarinic receptor stimulation are described in detail for endocrine beta cells and exocrine acinar cells separately. Here we describe morphological and functional criteria to separate these two cell types in situ in tissue slices and simultaneously measure their response to ACh stimulation on cytosolic Ca2+ oscillations [Ca2+]c in stimulatory glucose conditions. Our results show that both cell types respond to glucose directly in the concentration range compatible with the glucose transporters they express. The physiological ACh concentration increases the frequency of glucose stimulated [Ca2+]c oscillations in both cell types and synchronizes [Ca2+]c oscillations in acinar cells. The supraphysiological ACh concentration further increases the oscillation frequency on the level of individual beta cells, inhibits the synchronization between these cells, and abolishes oscillatory activity in acinar cells. We discuss possible mechanisms leading to the observed phenomena.
Ključne besede: pancreas tissue slices, acetylcholine, beta cell, acinar cell, Ca2+ oscillations
Objavljeno v DKUM: 14.10.2024; Ogledov: 0; Prenosov: 17
.pdf Celotno besedilo (2,41 MB)
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4.
In vitro disease models of the endocrine pancreas
Marko Milojević, Jan Rožanc, Jernej Vajda, Laura Činč Ćurić, Eva Paradiž, Andraž Stožer, Uroš Maver, Boštjan Vihar, 2021, pregledni znanstveni članek

Opis: The ethical constraints and shortcomings of animal models, combined with the demand to study disease pathogenesis under controlled conditions, are giving rise to a new field at the interface of tissue engineering and pathophysiology, which focuses on the development of in vitro models of disease. In vitro models are defined as synthetic experimental systems that contain living human cells and mimic tissue- and organ-level physiology in vitro by taking advantage of recent advances in tissue engineering and microfabrication. This review provides an overview of in vitro models and focuses specifically on in vitro disease models of the endocrine pancreas and diabetes. First, we briefly review the anatomy, physiology, and pathophysiology of the human pancreas, with an emphasis on islets of Langerhans and beta cell dysfunction. We then discuss different types of in vitro models and fundamental elements that should be considered when developing an in vitro disease model. Finally, we review the current state and breakthroughs in the field of pancreatic in vitro models and conclude with some challenges that need to be addressed in the future development of in vitro models.
Ključne besede: in vitro disease models, pancreas, islet of Langerhans, 3D cell culture, scaffolds, acute tissue slices
Objavljeno v DKUM: 01.10.2024; Ogledov: 0; Prenosov: 7
.pdf Celotno besedilo (4,42 MB)
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5.
Gas sensing system based on an all-fiber photothermal microcell
Matej Njegovec, Jure Javornik, Simon Pevec, Vedran Budinski, Tomaž Gregorec, Benjamin Lang, Manuel Tanzer, Alexander Bergmann, Denis Đonlagić, 2024, izvirni znanstveni članek

Opis: This article presents an all-fiber, miniature Fabry-Perot gas sensor based on photothermal absorption spectroscopy with a custom-made and cost-efficient interrogation system. The sensing gas microcell has a diameter of 125 μ m and a length of 1 mm, and allows for free gas exchange within the optical resonator through micromachined slits. High light intensity and confinement are ensured by delivering the excitation light directly into the gas microcell through a lead-in single-mode fiber. This enhances the photothermal effect and provides a short system response time. The interrogation system utilizes the modulation of an excitation laser diode with a fixed frequency while locking the probe laser onto the gas microcell’s quadrature point (QP) and observing the variations of the reflected optical power. To show the potential of the proposed system, nitrogen dioxide (NO2 was measured in dry air. Thereby, a limit of detection of 20 ppm could be achieved for 10 s integration time. Furthermore, the small dimensions of the sensor allow for improved dynamic performance with photothermal modulation frequencies as high as 7 kHz.
Ključne besede: Fabry–Perot interferometer, FPI, fiber gas cell, fiber gas sensor, nitrogen dioxide, NO2, photothermal effect
Objavljeno v DKUM: 29.08.2024; Ogledov: 96; Prenosov: 10
.pdf Celotno besedilo (15,77 MB)

6.
Glucose-stimulated calcium dynamics in beta cells from male C57BL/6J, C57BL/6N, and NMRI mice : a comparison of activation, activity, and deactivation properties in tissue slices
Viljem Pohorec, Lidija Križančić Bombek, Maša Skelin, Jurij Dolenšek, Andraž Stožer, 2022, izvirni znanstveni članek

Opis: Although mice are a very instrumental model in islet beta cell research, possible phenotypic differences between strains and substrains are largely neglected in the scientific community. In this study, we show important phenotypic differences in beta cell responses to glucose between C57BL/6J, C57BL/6N, and NMRI mice, i.e., the three most commonly used strains. High-resolution multicellular confocal imaging of beta cells in acute pancreas tissue slices was used to measure and quantitatively compare the calcium dynamics in response to a wide range of glucose concentrations. Strain- and substrain-specific features were found in all three phases of beta cell responses to glucose: a shift in the dose-response curve characterizing the delay to activation and deactivation in response to stimulus onset and termination, respectively, and distinct concentration-encoding principles during the plateau phase in terms of frequency, duration, and active time changes with increasing glucose concentrations. Our results underline the significance of carefully choosing and reporting the strain to enable comparison and increase reproducibility, emphasize the importance of analyzing a number of different beta cell physiological parameters characterizing the response to glucose, and provide a valuable standard for future studies on beta cell calcium dynamics in health and disease in tissue slices.
Ključne besede: beta cell, mouse models, calcium imaging, glucose-dependence, tissue slice
Objavljeno v DKUM: 15.07.2024; Ogledov: 148; Prenosov: 21
.pdf Celotno besedilo (4,45 MB)
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7.
pH-dependence of glucose- dependent activity of beta cell networks in acute mouse pancreatic tissue slice
Sandra Postić, Marko Gosak, Wen-Hao Tsai, Johannes Pfabe, Srdjan Sarikas, Andraž Stožer, Dean Korošak, Shi-Bing Yang, Marjan Rupnik, 2022, izvirni znanstveni članek

Opis: Extracellular pH has the potential to affect various aspects of the pancreatic beta cell function. To explain this effect, a number of mechanisms was proposed involving both extracellular and intracellular targets and pathways. Here, we focus on reassessing the influence of extracellular pH on glucose-dependent beta cell activation and collective activity in physiological conditions. To this end we employed mouse pancreatic tissue slices to perform high-temporally resolved functional imaging of cytosolic Ca2+ oscillations. We investigated the effect of either physiological H+ excess or depletion on the activation properties as well as on the collective activity of beta cell in an islet. Our results indicate that lowered pH invokes activation of a subset of beta cells in substimulatory glucose concentrations, enhances the average activity of beta cells, and alters the beta cell network properties in an islet. The enhanced average activity of beta cells was determined indirectly utilizing cytosolic Ca2+ imaging, while direct measuring of insulin secretion confirmed that this enhanced activity is accompanied by a higher insulin release. Furthermore, reduced functional connectivity and higher functional segregation at lower pH, both signs of a reduced intercellular communication, do not necessary result in an impaired insulin release.
Ključne besede: insulin secretion, membrane excitability, potassium channels, beta cell network, collective activity, calcium waves, pancreatic islets, pH-dependence
Objavljeno v DKUM: 01.07.2024; Ogledov: 149; Prenosov: 4
.pdf Celotno besedilo (7,45 MB)
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8.
The role of anaplerotic metabolism of glucose and glutamine in insulin secretion : a model approach
Vladimir Grubelnik, Jan Zmazek, Marko Gosak, Marko Marhl, 2024, izvirni znanstveni članek

Opis: We propose a detailed computational beta cell model that emphasizes the role of anaplerotic metabolism under glucose and glucose-glutamine stimulation. This model goes beyond the traditional focus on mitochondrial oxidative phosphorylation and ATP-sensitive K+ channels, highlighting the predominant generation of ATP from phosphoenolpyruvate in the vicinity of KATP channels. It also underlines the modulatory role of H2O2 as a signaling molecule in the first phase of glucose-stimulated insulin secretion. In the second phase, the model emphasizes the critical role of anaplerotic pathways, activated by glucose stimulation via pyruvate carboxylase and by glutamine via glutamate dehydrogenase. It particularly focuses on the production of NADPH and glutamate as key enhancers of insulin secretion. The predictions of the model are consistent with empirical data, highlighting the complex interplay of metabolic pathways and emphasizing the primary role of glucose and the facilitating role of glutamine in insulin secretion. By delineating these crucial metabolic pathways, the model provides valuable insights into potential therapeutic targets for diabetes.
Ključne besede: mathematical models, pancreatic beta cell, pyruvate-malate cycle
Objavljeno v DKUM: 06.06.2024; Ogledov: 162; Prenosov: 7
.pdf Celotno besedilo (5,53 MB)
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9.
Assessing different temporal scales of calcium dynamics in networks of beta cell populations
Jan Zmazek, Maša Skelin, Rene Markovič, Jurij Dolenšek, Marko Marhl, Andraž Stožer, Marko Gosak, 2021, izvirni znanstveni članek

Opis: Beta cells within the pancreatic islets of Langerhans respond to stimulation with coherent oscillations of membrane potential and intracellular calcium concentration that presumably drive the pulsatile exocytosis of insulin. Their rhythmic activity is multimodal, resulting from networked feedback interactions of various oscillatory subsystems, such as the glycolytic, mitochondrial, and electrical/calcium components.How these oscillatory modules interact and affect the collective cellular activity, which is a prerequisite for proper hormone release, is incompletely understood. In the present work, we combined advanced confocal Ca2+ imaging in fresh mouse pancreas tissue slices with time series analysis and network science approaches to unveil the glucosedependent characteristics of different oscillatory components on both the intra- and inter-cellular level. Our results reveal an interrelationship between the metabolically driven low-frequency component and the electrically driven high-frequency component, with the latter exhibiting the highest bursting rates around the peaks of the slow component and the lowest around the nadirs. Moreover, the activity, as well as the average synchronicity of the fast component, considerably increased with increasing stimulatory glucose concentration, whereas the stimulation level did not affect any of these parameters in the slow component domain. Remarkably, in both dynamical components, the average correlation decreased similarly with intercellular distance, which implies that intercellular communication affects the synchronicity of both types of oscillations. To explore the intra-islet synchronization patterns in more detail, we constructed functional connectivity maps. The subsequent comparison of network characteristics of different oscillatory components showed more locally clustered and segregated networks of fast oscillatory activity, while the slow oscillations were more global, resulting in several long-range connections and a more cohesive structure. Besides the structural differences, we found a relatively weak relationship between the fast and slow network layer, which suggests that different synchronization mechanisms shape the collective cellular activity in islets, a finding which has to be kept in mind in future studies employing different oscillations for constructing networks.
Ključne besede: islets of Langerhans, beta cell network, calcium oscillations, multimodal activity analysis, confocal imaging, functional connectivity, multiplex network
Objavljeno v DKUM: 06.06.2024; Ogledov: 171; Prenosov: 6
.pdf Celotno besedilo (9,40 MB)
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10.
Role of cAMP in double switch of glucagon secretion
Jan Zmazek, Vladimir Grubelnik, Rene Markovič, Marko Marhl, 2021, izvirni znanstveni članek

Opis: Glucose metabolism plays a crucial role in modulating glucagon secretion in pancreatic alpha cells. However, the downstream effects of glucose metabolism and the activated signaling pathways influencing glucagon granule exocytosis are still obscure. We developed a computational alpha cell model, implementing metabolic pathways of glucose and free fatty acids (FFA) catabolism and an intrinsically activated cAMP signaling pathway. According to the model predictions, increased catabolic activity is able to suppress the cAMP signaling pathway, reducing exocytosis in a Ca2+ -dependent and Ca2+ independent manner. The effect is synergistic to the pathway involving ATPdependent closure of KATP channels and consequent reduction of Ca2+. We analyze the contribution of each pathway to glucagon secretion and show that both play decisive roles, providing a kind of "secure double switch". The cAMP-driven signaling switch plays a dominant role, while the ATP-driven metabolic switch is less favored. The ratio is approximately 60:40, according to the most recent experimental evidence.
Ključne besede: pancreatic alpha cell, glucagon, cAMP, mathematical model, diabetes, cellular bioenergetics
Objavljeno v DKUM: 06.06.2024; Ogledov: 107; Prenosov: 11
.pdf Celotno besedilo (2,58 MB)
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